Anti-inflammation activity of Zhizi Jinhua Pills and overall quality consistency evaluation based on integrated HPLC, DSC and electrochemistry fingerprints.

Abstract

Zhizi Jinhua Pills (ZZJHP), a compound preparation composed of 8 traditional Chinese medicines (TCM), is widely used clinically to clearing heat, purging fire, cooling blood and detoxifying. However, the studies on its pharmacological activity and the determination of active compounds are relatively few. There is a lack of quality control methods that can reflect the effectiveness of the drug. The objective was to construct fingerprint profiles, conduct a spectrum-effect relationship study and establish an overall quality control method for ZZJHP through anti-inflammatory and redox activity studies. Firstly, anti-inflammatory activity was tested using the xylene-induced ear edema model in mice. Then, Five-wavelength fusion HPLC fingerprint, electrochemical fingerprint, and Differential scanning calorimetry (DSC) profiling were established to evaluate ZZJHP more comprehensively, where Euclidean quantified fingerprint method (EQFM) was proposed for the similarity assessment of these three fingerprints. Moreover, the spectrum-activity relationship of HPLC-FP and DSC-FP with electrochemical activity helped explore the active components or ranges in the fingerprint. Finally, integrated analysis of HPLC, DSC and electrochemistry were used for the quality screen of samples from different manufacturers. ZZJHP was found to significantly decrease the levels of both TNF-α and IL-6 in the mice. Qualitatively, the integrated similarity Sm of 21 samples were all greater than 0.9, indicating the great consistency in chemical composition. Quantitatively, 9 batches of samples were classified as Grade1∼4; 6 batches of samples were classified as Grade5∼7 due to higher PINT; 6 batches of samples were classified as Grade4∼5 due to lower PINT. EQFM can qualitatively and quantitatively characterize the fingerprint profile information from an overall perspective. This strategy will contribute to the quantitative characterization of TCM and promote the application of fingerprint technology in the phytopharmacy field.